Amino-sterol compounds and production thereof



United States Patent 3,424,747 AMINO-STEROL COMPOUNDS AND PRODUCTIONTHEREOF 3,424,747 Patented Jan. 28, 1969 "-OR or ,CR' F-CH R R being ahydrogen atom,an

-whilst R' is e f fl re e pnqr, 9y. Qup.de ed fr a carboxylic acidhaving a total of less than 6 carbon atoms,

Josef Schmitt, LHay-les-Roses, and Jacques J. Panouse,

Paris, France, assignors to Etablissements Clin-Byla, H 0 Paris, France,a company of France J N0 Drawing. Filed Sept. 9, 1960, Ser. No. 54,858 1Claims priority, apPlic835i0I110f8I1Ce, Sept. 12, 1959, 0 Cl. z z ,5 17m i m in which n is 2 or 3; and R is a hydrogen atom or --OR Int. Cl.C07c 173/10, 169/50; A61k 17/00 including the casein which there is anoxygen bridge between the carbon atoms 20 and 21 which carry the groupsThe present invention relates to a process for preparing R and R R is ahydrogen atom or. OR and is a hysterol compounds among which are newsterol compounds. drogen atom when R is -OR; The main Object of theinvention is to enrich the therapeutic armamentarium with compoundshaving aselective H action on the centralnervous system and which areparis =0; ticularly valuable as -neuroleptics and aggressiveness-in-. tthibitors, but are of low toxicity. p v 0H I A further object is toprovide a convenient route to a R4 is a hydrogen atom or a methyl group;whole class of sterol compounds which arejfor the most 7 H H part, new.f,

The invention relates to compounds having a primary, R5 is He, orsecondary or tertiary amino group (or functional derivative thereof suchas an amine salt, a quaternary ammoni- 25 0H; 0H 11m compound, an amidean imine) il'lflthe "P X represents either hydrogen atoms at positions4and 5, of a steroid substituted inter'alia at the 17-position either orotherwise there is a double b at position 4 but i y an y -e p 8 y Yestefnethel'eal the latter case Am can be only a pyrrolidine group or any y ham contalmng Y carboljl atoms and amine group of similar structure;Ami? is either (a) at having at least One oxygencontelmflg funetlonal.group primary amine group or an imine group of the general y y ketone,ester, ethereal Oxygen or ketal) or y formula -N=CH-Ar i which Ar is abenzene ring O Such P which may be substitutcd;'y(b) a lower aliphatic,aralie paftlclllafly the compounds correspond to the phatic orcycloaliphatic amino group, either unsubstituted fOHOWlIlg generalformula: W or having various substituents, and may be a heterocyclic CH3'35 ring containing a nitrogen atom; (c) an amido group R1 derived froma lower aliphatic acid and from a primary k -or secondary amine asdefined under (a). or (b) above; (d) is an amino group as defined under(a) and (b) above 11 13 16 in the form of a salt or ultimately convertedinto a qua- R 14 15 40 ternary ammonium compound, and comprising interalia 1 the compounds for which the above symbols have the m 2 meaningshown in Table 1 below. In that table and in the 3 5 7 remainder of thedescription tube means that the un- H''\*/% 6 corrected melting pointhas been determined using a H ,H} X capillary tube, (K) means that itwas determined using 7 H X B I a Kofler block, and (mien) that it wasdetermined in which R is a group having one of the general formulae.using a microscope.

TABLE I Posipound Position 17 (R1+R2) Posi- Positions Position 3Position 5 Melting Point, having on 11 tion 10 6 and X C For- 11 (5) R01) 11 R 08) 16 R Am H male I Ia -430-4333 ..H H9 0H3 H2 -N p .a H#132-133 (tube).

116-118 (tube It) -COCH3 -.H H: CH3 H2 ..N a -fl Hfl and K).

Ic '-0 0-011: ..H H: +0113 Hi -N a .0: ..Ha Egg? Id -'-0 0-01-13 ..H H,-CHa H: ..N a B Ha Egg?- Ie o ooH= ..H H1 -CHa H2 N 5 A4-5 136-138 (K).

If -o O-GH; ..H H: -CH: H2 -N(OH=) a H6 96-97 (tube).

Ig -o 0-0113 ..H H: CH: H: N B -HB 121-122 (tube).

m Ih c O-GH; ..H H2 H2 N 0 ..a 144-145 (tube).

TABLE ICont1nued Com- Posipound Position 17 (RH-R Posi- PositionsPosition 3 Position 5 Melting Point, having tion 11 tion 10 6 and X C.

For- R (fi) R (a) R RKB) 16 R Am H mula I Iaa COCH2OCOCH3 .H H: CH: H:-N=CH 6 B ..a H 13 132-134 (tube).

OH 6 Iap c o-omococm on -orn H, -wNH M- g Mn; Not detera mined.

OH H Iaq C O-CHzOCOCHa OH CH3 H; -wN=CH-@ w E -H 204-205 (tube).

Iar OHOH-CHaOH 1 ..H H: CH H: N B .....u --H 5 188-190 (tube).

1G8 CHOH-CH2011 .11 H2 CH: H: ...N 0: B H 5 254-256 (K).

. Iat CHOHCH:OH ..H H: CH3 H: N B .a A4-5 210-212 (K).

Iau CHQH-CHiOH H H: CH: H: ...N a [3 H B 238 (K),

I CHOH CH 011 H H OH H N O 8 H5 227 229 (K) av z 2 z a a CHOH OH OH H HCH H N N CH 8 Hfl 203-204 (K) Iaw 2 2 s n 3 a Ia: (JHOH-CH2OH ..H H2 CHaH: NH: 5 .....a HB 132-136 (K).

Iay CHOHCHzOH ..H H: CH3 H: --N=CH as B .....o: HB 156-158 (K).

110-115 In: CHCH3 H H: CH: Hz N(CH3)2 a B H 5 d I I 152-154 (K). O\ /OCHOH Iba (J H-(|3Hz ..H Ha CHa Ha .-.NH--CH2 a B Hfl 94-96 (K).

CHOH

sOH at 020.

Also included are the salts which the above-described compounds formwith such pharmaceutically acceptable mineral and organic acids ashydrochloric, formic, acetic, fumaric, maleic, citric and ascorbicacids, as well as in the case of the tertiary amines their quaternarycompounds, more especially the methiodides.

Prior to the present invention, only a small number of compounds ofFormula 1 above have been known. Thus all the compounds set out in TableI above are new and are comprised Within the scope of the invention.Among 10 I, both known and novel, can readily be prepared according toone or other of the following two methods, both of which use as startingmaterials ketones having the general Formula II:

in which R R R R, R and X are defined .above in connection with thegeneral Formula I and more especially those ketones for which thesymbols have the following meanings:

TABLE II Compound Position 17 (R +R Position Position Positions Positionhaving 11 10 6 and 16 5 Common name of Substance Formula II R (B) R (a)R R R (x-x) IIa COCH3 H H2 CHafi H2 A i-5 Progesterone. IIb... C OOH HH: CHafl H1 Ha Allopregnanedione. C O-CH3 H H2 CHaB H2 HB Pregnanedione.

-OH H H: -CH3B Hz Ha Androstanolone. OH H H: CHaB H2 A4-5 Testosterone.CO-CH2OH H H2 CHafl Hz A4-5 Desoxycorticosterone (DOC) -COOH1OH H HaOHZB Hz HB Hydro-DOC. OHOH-CHzOH H H2 -CH3B H1 HB (None). CO-CHgOCOCHs HH: CHQB Hz A4-5 Acetate of DOC (DOCA). CO-CHzOCOCHa H H: CHaB H2 H5Hydro-DOOA. IIL COCH;OH 0H =0 OHafi H2 134-5 Cortisone. 1n co-omococm 0Hgi oHm H2 44-5 Hydrocortisone acetate.

The first method consists in treating a ketone having the generalFormula II with an amine, preferably a secondary amine, and with formicacid: these are used simultaneously if the ketone used lacks a doublebond between positions 4 and as well as a free ketone group in the sidechain attached to the 17-carbon atom. Otherwise they can be used oneafter the other, that is to say with the formation of an intermediateenamine which is then reduced by the formic acid. The process in thislatter form is of interest because it can be applied to polyketosteroidsand to those with a double bond between the 4 and 5 positions. In thiscase, however, it is limited to amines which readily give with ketoneswhat is conveniently called an enamine; in particular, to pyrrolidine.It is also limited to compounds having the general Formula H which havea methyl group in position (R =CH and which are free from a hydroxylgroup at position 6. In the form of its two variants the process ingeneral leads mainly to steroids having an amino group orientatedequatorially at position 3 if ring A is saturated, or quasi-equatoriallywhen there is a double bond at position 4.

Certain of the intermediate enamines having the general Formula IIIgiven below are new compounds, more particularly the3-pyrrolidinyl-21-hydroxypregna-3,5-diene--one and3-pyrrolidinyl-21hydroxy-Sfl-pregn-3-ene- 20-one.

In the general Formula III R R R R and Am are as defined in connectionwith general Formula I, while Y represents either -H at position 5 and Hor at position 6, or otherwise a double bond between the 5 and 6positions with either H or -CH at position 6.

R1 H3O 2mm III The second method enables the preparation, usually as themain product, of steroid amines having an amino group, having the [iconfiguration at position 3, while the 30:. isomers may occasionally beisolated in substantial amounts from the reaction mixture. It comprisestreating ketones of the general Formula II with ammonia or a primary orsecondary amine and with hydrogen in the presence of suitablehydrogenation catalysts.

The Am group can have any of the meanings given to it in connection withFormula I above. Also the ketosteroids II used as starting materials mayhave one or more substituents which are inert towards hydrogen under theconditions of hydrogenation.

Basically the groups R and R (Formula I) at position 17 in thecyclopentanoperhydrophenanthrene nucleus remain unchanged, whatever themode of operation used, but secondary reactions may nevertheless occur.Thus hydrolysis of ester and other groups known to be extremely labilewhen in contact with aqueous acid or alkali, can occur unexpectedly. Inthe same way esterification of hydroxyl groups present is sometimesobserved, more especially the formation of cyclic orthoformic esterswhen a compound having the general Formula II having a glycol groupingat positions 20 and 21 is in contact in the reaction mixture with aformamide, which latter has been generated spontaneously during thecourse of the reaction or has been added as a source of formic acid.

To determine the spatial arrangement of the compounds described herein,the applicants have made the 3-paratoluenesulphonates (tosylates) of the3-hydroxy-steroids and then submitted them to aminolysis, the3-hydroxysteroids and the amines being chosen so as to give 3- aminocompounds of planar configuration identical with that of the compoundshere described. Comparison has then been made, taking into account thewell-known fact that aminolysis is accompanied by inversion ofconfiguration (SN2 type of reaction; see for example: Pierce andco-workers, J. Chem. Soc., 1955, page 694; Shoppee and co-workers, J.Chem. Soc., 1956, page 1649; Sauers, J. Amer. Chem. Soc., 1958, volume80, page 4721).

Thus. in the 5 aH series, by reacting pyrrolidine under slight pressureand at about C. with tosylated 3ehydroxy-Sea-pregnane-ZO-one (Ruff andReichstein, Helv. Chim. Acta, 1951, volume 34, page 70) made by reactingthe latter with tosyl chloride, 3u-pyrrolidinyl-5a-pregnane- 20-one(Formula IV) is obtained (M.P. (K); -106-108 0, (tube): 104-1'06 C.),identified with the compound Id, isolated as a by-product when hydrogenand pyrrolidine react with allopregnanedione 11b in the presence ofpalladium supported upon charcoal. The melting point and mixed meltingpoint are identical. In addition the inthe principal product obtained bythe action of hydrogen and pyrrolidine on allopregnanedione 11b isidentical with that obtained on reduction by formic acid of thepyrrolidine enamine corresponding to 11b. The structure 10 attributed tothem is thus certainly correct.

Actually the presence of a keto group at position 20, allo to position5, is equally well confirmed for Is as for Id, by examination of theinfrared spectrum (R. Norman Jones, F. Herling and E. Katzenellebogen,J. Amer. Chem. Soc., 1955, volume 77, page 651). Since 10 and Id areisomers they can only differ with respect to the configuration aboutcarbon atom 3. The pyrrolidine group in Ic can then only be 33 and henceequatorial.

In the same way, in the normal series (S BH), hydro-4,SB-desoxycorticosterone (Hg) has been reduced by sodium borohydride. Theconfiguration 3a-OH, 20/8-OH is ascribed with complete certainty to thetriol obtained in the reaction. Detailed examination of the infraredspectrum does in fact confirm this stereo-configuration, which was apriori foreseen on the basis of the analogous reactions described in theliterature (see for example: Nussbaum, J. Amer. Chem. Soc., 1959, volume81, page 1228; Wheeler and Mateos, Chem. and Ind., 1957, page 395). Thetriol is then treated with acetone in the presence of anhydrous coppersulphate (as described by Reichstein, United States patent specificationNo. 2,423,517). In this way the hydroxyl groups of the ot-glycol portionof the side chain are selectively blocked as a ketal. The latter is thentreated with para-toluenesulphonyl chloride and the free hydroxyl groupat position 3 is thereby esterified. The ketal ester is not isolated.The very labile ketal group is hydrolysed during attempts atrecrystallisation and actually the ester of the glycol is obtained. Thelatter is treated with pyrrolidine at about 120 C. under slightpressure. Aminolysis of the tosylate brings about inversion ofconfiguration at position 3 and a satisfactory yield 9 of3fi-pyrrolidinyl-Sit-pregnane-ZOBJl-diol (Iar) is isolated, M.P. 187191C.:

CHQOH HOCH This aminated sterol diol with a 3B axial configuration hasproved to be identical with the compound obtained by the action ofpyrrolidine and hydrogen, in the presence of palladium supported uponcharcoal, on 205,21-dihydroxy- 8-pregnane-3-one (Hit). The melting pointand mixed melting point are identical and in addition the infraredspectra are superimposable.

On the other hand, the product obtained by reducing the same startingketone III: with pyrrolidine and formic acid is an aminated diolisomeric with the preceding one, but differentiated sharply therefrom byits infra-red spectrum, by its melting point ((K): 254-256 C.), and bythe depressed mixed melting point obtained from the two substances. Thusthe diol produced by reduction with formic acid is given the formulaIas, 3u-pyrrolidinyl-5ppregnane-20 ,8,2l-diol, with a 30: amino group,which is thus equatorial.

To ascertain the structure of the amines produced by reduction withformic acid of the pyrrolidine enamine derivatives of 3-ketosteroidshaving a double bond in the 4-5 position, the3B-pyrrolidinyl-pregna-x-ene-20-one, obtained by the action of formicacid on the pyrrolidine enarnine of progesterone, is treated withhydrogen in the presence of palladium. One molecule of hydrogen is takenup and 3fi-pyrrolidinyl-Sa-pregnane-ZO-one, Ic, whose configuration isknown with certainty (see above), is isolated as the sole reactionproduct. This enables an unequivocal assignment of the 3-{3configuration to the amino group of the unsaturated amine, since thecentre of asymmetry at C3 is not able to change during thehydrogenation.

On the other hand, the double bond present in the unsaturated aminescannot be elsewhere than between positions 4 and 5. Actually, accordingthe method of preparation the double bond could only be between thecarbon atoms 4 and 5 or alternatively between 5 and 6. But thedifferences in the molecular rotation between these unsaturated aminesand the steroids having the nuclei A and B saturated (50c series) isbetween +55 and +65. It is therefore of the same order of magnitude asthe difference in the molecular rotation between3fi-benzylaminocholest-4-ene and 3B-benzylamino-5u-cholestane (thedifference in molecular rotation lies between +18 and +32) and is in nowise comparable with the difference in molecular rotation between the3B-aminosteroids having a double bond between the 5 and 6 positions andthe corresponding reduced derivatives (Su-H), which is between 132 and245 (cf. Janot, Cave and Goutarel, Bull. Soc. Chim., France, 1959, page836).

Study of infra-red spectra has made a useful contribution towardsdetermining the structure of the aminosteroids prepared according to thepresent invention. Thus a certain number of bands characteristic of eachtype of spatial isomerism at positions 3 and 5 could be adduced asevidence, but most of the bands vary according to the nature of theamino group and can no longer be recognised in the presence in themolecule of functional groups with a very marked tendency to associate(especially bydroxyl groups) or again if the sterol amine isinvestigated in the form of a salt. Subject to these reservations, ithas been noted that primary amines give rise to absorption which is notvery intense, but which is sharp, over the region 6.25-6.30 r(deformation of N-H bond) only for those sterols aminated at the3a-position, whilst those sterols aminated at the 3fl-position havelittle or no absorption in this region. Furthermore, there has beennoted in the 14.70/L region a band which is characteristic only of thecompounds having an axial primary amino group and which is completelyabsent from the equatorial derivatives.

For the tertiary amines:

(I) In the region 7.0-7.6 (absorption doubtless due to vibrations of l adoublet is always observed with those compounds having the amino groupequatorial and a hydrogen atom axial, whilst if the amino group is axialand a hydrogen atom is equatorial there is a triplet. It is evident thatspectra measured in suspensions in liquid par-afiin cannot give any useful information in this region.

(II) Most of the steroids with a tertiary amino group absorb in theregion 11.35-11.60 the equatorial amines have a band at 1136-1138,,while the corresponding axial amines have a band between 11.50 and11.60,u.

Furthermore, the study of the infra-red spectra sometimes gives (moreespecially when hydroxyl groups are absent) very precise informationconcerning the configuration at position 5 (whether a or B) for theZO-keto-steroids, as the presence of a functional amino group'atposition 3 does not seem in the least to disturb the characteristicbands of normal or allo ketones over the band between 1300 cm.- and 900cmr It is to be noted that the infra-red spectra have been recorded witha double-beam Baird spectrograph having sodium chloride prisms, usingcompensated solutions (chloroform, methylene dichloride, carbondisulphide or carbon tetrachloride) or a suspension in liquid parafiinexcept that certain steroids melting below C. have sometimes beenexamined as a thin layer, after melting and resolidification on arocksalt plate (microcell).

The compounds set forth in Table I have been submitted to many testsaffording evidence about their pharmacodynamic properties, and inparticular as to their action on the central nervous system. Among thetests may be mentioned: toxicity tests on mice, rats and cats;impairment of balance, mainfestations of catatonia; degree of anorexia;modification of aggressiveness (rat with respect to rat, cat with regardto mouse), effect on spontaneous movement or that induced by benzedrine;antagonism to the catatonia-inducing effect of perchlorperazine, andeffect on the behaviour of conditioned animals.

These compounds have been administered as the base or as salts, eitherparenterally or orally.

In general the compounds Io, Ip, Iq and lam, having an amino group inthe 3-position, have shown themselves to be sedatives. The compound Ioshowed itself to have exceptional properties. It has moderate toxicity(DL 50 IP mg./kg., DL 50 oral 1g./kg.).

Administered intraperitonally it allays aggressiveness in rats at adosage of 15 mg./kg. and aggressiveness in cats at 2.5 mg./kg., and thisaction persists for 24 hours. When administered to cats in doses of 10and 20 mg./kg., the sedative action lasts several days. At these dosagesthe product does not affect either the arterial blood pressure or thecardiac rhythm. Doses of 50 mg./kg. must be used to affect the motorfunctions of the rat or the balance of the mouse. With high doses (100mg./kg.) catatonia is not induced in mice.

This product 10 has a synergistic effect when used with varioussedatives or tranquillisers. If a rat is simultaneouly given anineffective dosage (1 mg./kg.) of this compound together with an equallyineffective dosage of acepromazine (1 mg./kg.) or of chlorpromazine, itproduces a powerful sedative effect.

Another especially active product is 3a-pyrrolidinyl-21-hydroxy-SB-pregnane-ZO-one (Iaf). Slightly more toxic than I0, it hasshown sedative properties with respect to aggressiveness in rats atdosage between 50 and 75 mg./kg. administered intraperitonally. Given tocats a sedative effect is observed from mg./ kg. and at dosages of to 20mg./kg. this effect can last for several days. At such dosages thiscompound neither modifies arterial pressure nor cardiac rhythm andleaves the motor conduction and those functions for preserving balancepractically unchanged.

Other compounds of interest are: the Iao compound, which is abenzylideneamine derivative of hydrodesoxycorticosterone acetate IIj; Ib(the 3a-pyrrolidinyl derivative of SB-pregnane-ZO-one); Iac (the3a-pyrrolidinyl derivative of Sfi-pregnane-ZOfi-ol); Ias (the3a-pyrrolidinyl derivative of 5 8-pregnane-20B,21-diol), and thecompounds Iar (the 3,3-pyrrolidinyl derivative of SB-pregnane-20B,21-diol) and Iau (the 3u-piperidinyl derivative of5fl-pregnane-20fi,21-diol) The first method which can be used for makingaminosteroids having the general Formula I consists in simultaneouslytreating ketones having the general Formula II with an amine, preferablya secondary amine, and formic acid, under the conditions for carryingout the Lenckart reaction, which has already been applied to certainsteroids by R. R. Sauers (J. Amer. Chem. Soc., 1958, volume 80, page4721) for the preparation of Sfi-monoalkylamino and 3fi-dialkylaminocholestanes, free from oxygen-containing substituents on the 17-carbonatom or in the side chain.

The applicants have found that if there are hydroxyl groups in thesteroid used as starting material, these are not in general esterifiedby formic acid even though it be added in an amount in marked excess ofthat theoretically required.

As an exception it has been observed that formation of a cyclicorthoformate occurs when a compound of the general Formula II having anu-glycol grouping at positions 20 and 21 is in contact, in the reactionmixture, with a formamide, the latter having arisen spontaneously in thecourse of the reaction or having been added as a source of formic acidand amine under the working conditions employed.

' Moreover, the applicants have found that not only are purer productsobtained but that these products are obtained more rapidly if thereaction mixture is not finally treated with hydrochloric acid toproduce a hydrochloride which has to be isolated in a solid state beforethe base is liberated, but rather if one dilutes the reaction mixturewith water and then makes the diluted reaction mixture alkaline so thatthe base passes into the organic phase, from which it can then beisolated in a state of great purity by crystallisation.

The following examples illustrates the first procedure in its first form(use of amine and formic acid together).

EXAMPLE 1 3ee-piperidinyl-SB-pregnane-20 3,2l-diol (Ian) To 5 g. of203,21-dihydroXy-5B-pregnane-3-one (IIh) (M.P. (K): 163 C.) are added 6ml. of piperidine. The mixture is heated to 80 C. in a 25 cc. flaskfitted with a reflux condenser, to effect solution. 4.5 mls. of formicacid are then added dropwise. A brisk reaction takes place. Thetemperature is then raised in an oil bath which is at 165 C. to 200 C.,more specifically to about 175 C. and maintained thereat for 7 to 10hours.

After cooling, the product is diluted with 500 cc. of water andacidified, if necessary, with formic acid. It is then extracted thricewith ether to remove the neutral fraction. The aqueous solution is thenmade alkaline with dilute sodium carbonate solution. A little ether isadded and the free amine crystallises. The solvent is allowed toevaporate, the crystals dried, washed with distilled water and thendried in vacuo. The product is 3u-piperidinyl- 125fi-pregnane-20fi,21-diol, 4.7 g.; M.P. (K): 200-220 C.Recrystallisation from n-propanol raises the melting point to 238 C.(K).

EXAMPLES 2-4 Example 1 is repeated respectively replacing the piperidineby morpholine, pyrrolidine and N-methyl piperazine. The followingcompounds are obtained in the same way:

3a-morpholinyl-Sfl-pregnane-Z05,21-diol (Iav), melting point afterrecrystallisation from methanol: M.P. 227- 229" C. (K); ultimately thecompound decomposed on the block.

3a-pyrrolidinyl-5,8-pregnane-20fl,2l-diol (Ias) melting point afterrecrystallisation from methanol: M.P. 254- 256 C. (K); ultimately theproduct decomposed on the block.

305 (4-methyl-1-piperazinyl)-5B-pregnane-20B,2l-diol (Iaw), meltingpoint after recrystallisation from ethyl acetate: M.P. 203 204 C. (K);ultimately the product decomposed on the block.

EXAMPLE 5 Preparation of the orthoformate of 3a-benZylamino-5/3-pregnane-20;3,21-diol (Iba) To 1.7 g. of20,21-dihydroxy-5-pregnane-3-one (IIh), 0.005 mol. are added 2.2 ml. ofbenzylamine (0.02 mol), followed by 1.5 ml. of formic acid (0.04 mol).The mixture is heated in a sealed tube, in an oil bath at 170- 180 C.for 10.5 hours; it becomes homogeneous. After cooling, the amber-yellowproduct is dissolved in a little ethyl alcohol, then 10 ml. ofconcentrated hydrochloric acid added. The mixture is boiled under refluxto hydrolyse the formamide formed as an intermediate. The hydrochorideof the amino-steroid formed is precipitated with water, dried, suspendedin chloroform and the amine displaced by adding ammonia. It passes intothe chloroform whence it is isolated as an oil after drying with calcuimchloride and evaporation of the solvent.

The orthoformate of 3a-benzylamino-SB-pregnane-ZO/S, 21-diol (Iba) isobtained crystalline after dissolving the oil in acetone andv slowlyevaporating the solvent. Yield: 1.08 g., M.P. (K): 93-96 C.

A pure sample for analysis of Iba is obtained on recrystallisation from60% aqueous acetone, M.P. 9496 C. (K). The hydrochloride of the baseIba, recrystallised from methanol, melts at 254260 C. (K).

The infra-red spectrum of the molten base shows the absence of compoundswith the 0:0 group, while a broad band towards 1040 cm.- may very wellbe bands due to the CO-C of the glycol orthoformate (cf. Bodenbenner,Annalen der Chemie, 1959, volume 623, page 183).

EXAMPLE 6 Preparation of the orthoformate of 3a-dimethylamino-5fl-pregnane-20,B,2l-diol (Iaz) from 205,21-dihydroxy-5fl-pregnane-3-one (IIh), dimethylformamide and formic acid 1.23 g. of205,21-dihydroxy-5fl-pregnane-3-one (IIh) is dissolved in 1.05 ml. ofdimethyformamide. 0.54 ml. of

formic acid is added dropwise to this solution. A copious whiteprecipitate is formed. The mixture is heated under reflux for 6 hours onan oil bath which is between and 190 C.,' more especially between and C.After cooling the partly crystallised reaction product is dissolved inwater and filtered to remove a small amount of an insoluble productwhich is washed with formic acid. The filtrate is extracted with etherto extract the non-basic material. The aqueous solution is then madealkaline and the liberated amine extracted with a little ether. Thesolvent is then allowed to evaporate and the dirty-grey product washedwith water and dried in vacuo,

weight 0.371 g.

After treating with isopropyl ether, an amino ester is isolated incrystalline form and purified by recrystallisation from ethyl acetate.The pure product has a double 13 melting point, firstly at 110-115 C.and then at 152- 154 C. (K). Its empirical formula corresponds with thatof a monoformate of 3u-dimethylamino-S/i-pregnane- 20/9,21-diol. Byanalogy with Example 5, however, it is more likely that it is theorthoformate of 3a-dimethylamino-Sfl-pregnane-ZOBJl-diol (Iaz).

When the secondary amine used can readily give an enamine having thegeneral Formula IH with the sterol ketone employed, it is preferable toeffect the reductive amination in two stages:

(a) Enamination;

(b) Formic acid reduction of the enamine.

Actually the steroid employed as starting material may contain aplurality of keto groups, in particular one in position 20, without anamino group being introduced anywhere other than at position 3.

In fact it it well known from the work of Herr, Heyl and theirco-workers (see particularly J. Am. Chem. Soc., 1953, volume 75, page1918; 1953, volume 75, page 5927; 1956, volume 78, page 430; 1955,volume 77, page 488; 1956, volume 78, page 500; French patentspecification No. 1,098,526; German patent specification No. 1,011,-419; United States patent specifications Nos. 2,733,072 and 2,781,368,and British patent specifications Nos. 740,- 568, 756,400, 763,835,763,836 and 779,001) that enamination of sterol polyketones takes placesolely at position 3, subject only to the rare exceptions pointed out bythese authors. 4

Hence it is unnecessary, in this embodiment of the invention,selectively to block carbonyl groups other that the carbonyl group atposition 3 before carrying out the reductive enamination. This is a verygreat advantage in comparison with the classical Leuckart reaction,because very much more readily accessible starting materials can beused.

Moreover, it is a well-known fact that unsaturated 04-5- ketones cannotbe converted into amines by means of the Leuckart reaction (OrganicReactions, volume 5, page 310).

Now the applicants have found that by reductive amination in two stages,steroids aminated at position 3 can be obtained without difficulty fromthe corresponding 3-ketones with a double bond between the 4 and 5positions. The formyl reduction of an intermediate enamine having onedouble bond at position 3 and another at position 5 results inunsaturated amines having a double bond at position 4; this can bededuced from calculations of the molecular rotation (see above).

In order to carry out the enamination, which is the first stage of theprocess of the invention, it is possible to proceed as described in theliterature, in particular by Herr and Heyl (loc. cit.). However, theapplicants have found that a better yield of enamines, particularly ofthose derived from ketones without a double bond at the 4 position, isobtained if the secondary amine is added to a benzene solution of the3-ketosteroid which has previously been brought to the boil underreflux.

With regard to the reduction of the enamines, this is effected by theaddition of formic acid to a solution of the enamine in an organicsolvent. Usually the reduction takes place in the cold, but it is oftenconvenient to heat somewhat, at least to initiate the reaction. Choiceof solvent is important, since with some the desired reduction does nottake place. In general aromatic hydrocarbons such as benzene, toluene,or xylene may be used; these are mentioned by way of example only.Though it is sometimes desirable to isolate the intermediate enamineprior to effecting reduction thereof, it is often preferable to reduce afreshly-prepared benzene solution of the enamine, made as describedabove, by means of formic acid. This merely necessitates use of anexcess of acid to neutralise the amine, which latter is always presentin considerable excess.

On the other hand, it is sometimes useful, in order to avoid as far aspossible resinification of the intermediate enamine (which is alwaysvery unstable when it is prepared in the hot, even under nitrogen) toeffect the operation in the cold using a very slight excess of secondaryamine, more particularly pyrrolidine, preferably in a polar organicsolvent having a fairly low boiling point, more especially methanol. Thequantity of solvent employed is not critical for successful Working andit is not necessary that all the steroid be dissolved therein. Additionof pyrrolidine brings about solution, followed by immediaterecrystallisation of the enamine which is only slightly soluble in thereaction mixture. It is then suflicient to evaporate the solvent and theslight excess of pyrrolidine at a very low pressure. It should be notedthat in a nonpolar solvent, such as benzene, the reaction does not takeplace (or is reversible) in the cold, except in the presence ofdehydrating agents such as calcium carbide.

The crude enamine is then redissolved in benzene and treated with formicacid in accordance with the procedure already described.

Several typical examples which follow illustrate the several methods ofoperation for carrying out the reductive amination of the invention intwo distinct stages.

EXAMPLE 7 Preparation of Kilt-pyrrolidinyl 3 5 androst-4-ene-l7/3-ol(Iy) via the pyrrolidinyl enamine derived from testosterone.

A flask, fitted with Soxhlet apparatus containing a thim'ble filled withpowdered calcium carbide, is charged with 5.76 g. of testosterone (I10),6 ml. of pyrrolidine, and ml. of benzene. After an hours boiling underreflux, no more bubbles are formed inside the calcium carbide, and theenamination is regarded as completed. Heating is discontinued, theSoxhlet apparatus is replaced .by a reflux condenser, and 4 ml. offormic acid is then added dropwise to the reaction mixture. A violentreaction takes place. There is a sharp evolution of gas, and anorange-coloured oil is formed, which floats on the surface of thebenzene. When the reaction slackens, the mixture si heated under reflux.After 3 hours it is allowed to cool. The lower, orange-coloured, layercrystallises spontaneously. After filtering, the long needles are suckeddry and washed with benzene. A little oily product passes into thefiltrate. The crystalline product is dried in vacuo; yield 5.623 g.(having M.P. (K): l60-l80 C.: pasty state). The product is the stillimpure formate of 3 6-pyrrolidinyl-androst-4-ene-175-01.Recrystallisation from isopropanol and dioxane gives it the form of paleyellow crystals (M.P. (K): -180 C. decomp.)

The benzene solution and the oily product which passes through thefilter are treated with dilute formic acid. The benzene layer containingneutral compounds is removed in a separating funnel. The aqueous layeris then made alkaline with sodium carbonate solution and the liberatedamine is then extracted with benzene, and the benzene solutions driedover sodium sulphate. After removing the solvent in vacuo, the aminecrystallises spontaneously. After drying the yield is 1.042 g. havingM.P. 184-186 C. (K).

Recrystallisation from ethyl acetate gives pure3,8-pyrrolidinyl-androst-4-ene-17,8-01 (Iy) M.P. (K) l96203 C. (slightdecomp.); [a] =+23 (c.=1% in chloroform).

EXAMPLE 8 Preparation of 3,8-pyrrolidinyl-pregn-4-ene-20 one (Ie) viathe enamine pyrrolidinyl derivative of progesterone A three-necked flaskis fitted with a tube for introducing nitrogen, and in the flask isprepared a benzene (100 ml.) solution of 18.35 g. of the pyrrolidinylenamine of progesterone (M.P. l92-l94 C. (K)). This compound is obtainedby the action of pyrrolidine on progesterone in solution in methanol asdescribed by Heyl 15 & Herr (J. Am. Chem. Soc. 1956, volume 78, page434).

2.5 ml. of formic acid are added drop by drop, to this solution. Avigorous reaction ensues. The mixture is then heated for 30 minutesunder reflux. After cooling, a further 2.5 ml. of formic acid is added,and then the mixture is evaporated to dryness in vacuo. The brown pasteobtained is dissolved in water, and the solution is thrice extracted toremove neutral compounds, consisting of slightly impure progesterone(weight 1.06 g.).

The aqueous solution is then made alkaline with sodium carbonate; avoluminous white precipitate of amine is formed. A little ether is addedand the liquid filtered. The greater part of the amine, which is onlyslightly soluble in this solvent collects upon the filter while thefiltered ether solution, separated from the aqueous phase by decantationyields, on evaporation, a further crop of crystals, the purity of whichis substantially the same as that of the first crop. Total weight afterdrying 11.856 g., M.P. (K) 128-134 C.

An analytically pure specimen of 3p-pyrrolidinyl-pregn- 4-ene-20-one(Ie) obtained by recrystallisation of the preceding product from ethylacetate had M.P. (K) 136-l38 C., [a] :+81.7 (c.:l.0% in dioxane), [a]=+99.6 (c.=1.0% in chloroform).

The above amino-ketone has been characterised by the followingcrystalline derivatives:

(a) methiodide, C H ONI, formed by reaction with methyl iodide M.P. (K)=237-242 C. (decomp.)

(b) the corresponding alcohol 3fi-pregn-4-ene-20B- o1 (Ind), obtained bythe action of lithium aluminum iodide, followed 'by recrystallisationfrom ethyl acetate, M.P. (K) l74177 C. [a] =+10.9 (c.=1.0% in dioxane).

EXAMPLE 9 Preparation of 3 8 pyrrolidinyl-2l-hydroxy-pregn-4-ene- 20-one(Iag) via the pyrrolidinyl enamine derivative of desoxy corticosterone(a) Preparation of the intermediate enamine, a new compound, by theaction of pyrrolidine either on desoxy corticosterone (111') or on itsacetate (Hi).

15 g. of desoxy corticosterone acetate are dissolved in 100 cc. ofmethanol. Then, whilst passing a stream of dry nitrogen, 6 ml. ofpyrrolidine is added all at once. The mxiture is boiled for 8 minutesand then allowed to cool under an atmosphere of nitrogen. The enamine or3-pyrrolidinyl-21-hydroxy pregn 3,5 diene-20-one crysallises out. It isfiltered off, sucred dry and then dried. Yield: 13.343 g. M.P. (K)163-l65 C.

(c.=1.0% in chloroform).

(b) Formic acid reduction of the enamine.

13.971 g. of the enamine prepared as described above are dissolved in 35ml. of boiling dry benzene. The solution is partly cooled and 2.1 ml. offormic acid added. Bubbles of gas are evolved and the mixture becomesorange-coloured. Two layers form, then the colour becomes paler and themixture becomes homogeneous.

It is then boiled for 55 minutes under reflux and the benzene removed invacuo. The residue crystallises spontaneously.

Recrystallisation from ethyl acetate gives3,B-pyrrolidinyl-21-hydroxy-pregn-4-ene-20-one (lag), M.P. (K): l74182C. yield 8.510 g. Further recrystallisation from methanol raises theM.P. to 184-l86 C. (K) [cc] =+89.5 (c.=l.0% in chloroform).

This hydroxy-amino ketone has been characterised thus:

(a) by its acetate, or 3,3-pyrrolidinyl-2l-acetoxypregn-4-ene-20-one(Ial), made 'by the action of acetic anhydride in the cold andrecrystallisation from ether; M.P. (K) 113118 C.;

(b) by preparation of the diol: 3fi-pyrrolidiny1-pregn- 4-ene-20,B, 21diol (Iat), by reduction with sodium borohydride, followed byrecrystallisation from methanol; M.P. (K) 210-212 C.

EXAMPLE 10 Preparation of 3a pyrrolidinyl 5B pregnane 20 one (Ib).

A flask, fitted with a Soxhlet apparatus containing a thirn'ble filledwith calcium carbide, and containing 6 g. of 5fi-pregnane-3,20-dione(IIc; M.P. ll2ll4) and 600 cc. dry benzene is heated to reflux. After 2hours, 6.5 ml. pyrrolidine is added all at once and the mixture boiledovernight. The Soxhlet apparatus is then removed and 10 ml. of formicacid added all at once. There is a copious evolution of fumes, and twolayers are formed. A current of nitrogen is passed through the still hotsolution, which latter is mechanically stirred for one hour. The mixtureis then concentrated in vacuo on a water bath, to a quarter of itsinitial volume, then diluted with 250 ml. of water and the neutralmaterial extracted with ether. The aqueous layer is then made alkalinewith a 20% solution of sodium carbonate. The free sterol amine isextracted with benzene, the benzene solution washed with water and driedover sodium sulphate. After filtering and then evaporating in vacuo,there remains an oily residue (yield 6.7 g.) which crystallisesspontaneously. The 3a-pyrrolidinyl-Sfi pregnane-ZOfi-one (Ib) ispurified by recrystallisation from 25 ml. of petroleum ether, weight=3g. (yield 43%); M.P. (K) 116-118 C.

An analytical sample is obtained after a further crystallisation frompetrol ether. The melting point is unchanged.

The presence of the keto group in the above ketone is shown by reductionwith sodium borohydride. There is thus obtained3a-pyrrolidinyl-5,8-pregnane-20/3-ol (lac); after recrystallisation frompetroleum ether M.P. 152- 156 C. (double M.P.).

EXAMPLE 11 Preparation of 3/3-pyrrolidinyl-5aregnane-ZO-one (10) Theprocedure is exactly as in Example 10, using as starting materialallopregnadione (IIb), and the intermediate enamine is not isolated.After recrystallisation from ethyl acetate the3l3-pyrrolidinyl-5a-pregnane-20 one (Ic) melts at 128130 C. (K).

EXAMPLE 12 Preparation of 3a pyrrolidinyl 21 hydroxy 5B pregnane-ZO-one(Inf).

(A) Via the pyrrolidinyl enamine of 2l-hydroxy- 5fl-pregnane-3,20-dione,a new compound.

5 g. of 21-hydroxy-5;8-pregnane-3,20-dione (Hg) is dissolved in 50 m1.of boiling methanol contained in a 250 cc. flask having a ground glassneck. The solution is cooled to room temperature, and 1.5 ml. ofpyrrolidine is added, whilst a current of dry nitrogen to the supersaturated solution thus obtained. The temperature is kept down bycooling with water. After some minutes, the enamine crystallises outfrom the solution, which has become a straw yellow colour. The mixtureis evaporated at 0.01 mn. of mercury, cooling being avoided by placingthe reaction flask in a bath of cold water when the methanol does notcome off. The white crystalline residue obtained is3-pyrrolidiny1-21-hydroxy-5BH-pregn-3-ene- 20-one, analytically pure,M.P. (K) 148-153 C. (decomp.)

This enamine is dissolved in 50 ml. of cold dry benzene. Then 1.5 ml. offormic acid is added, very slowly, whilst maintaining an atmosphere ofnitrogen and with mechanical stirring. Two layers are formed. Afterstirring for 20 minutes at room temperature, the aqueous layer becomesopaque. After 1 hour, a further 1 ml. of formic acid is added, and thetwo layers form again. Finally the mixture is warmed at 6065 C. for 5minutes, then allowed to return to the temperature of the laboratory,diluted with water, the phases separated and the aqueous layer extractedwith ether, in order to remove neutral compounds. The aqueous layer isthen made alkaline with an aqueous 20% sodium carbonate solution. Thefree amine crystallises. After allowing to stand it is dried, thenwashed with water and again dried. The crude 3m pyrrolidinyl 21 hydroxy5B pregnane 2O one (Iaf) weighs 4.65 g. and melts at 142-148 C. A singlerecrystallisation from boiling ethyl acetate gives it pure for analysis:weight 2.47 g. M.P. (K) l57159' C. ['a] "=+94.4 (c.=1. in chloroform). Afurther crystallisation from methanol did not alter these constants.

In another experiment the yields were better, (less neutral compoundswere isolated), as a result of a slightly longer heating time during theformic acid reduction of the enamine.

The above hydroxyamino ketone (Iaf) has been characterised by thefollowing derivatives:

(a) Acid fumarate of 1:1 prepared in solution in isopropyl alcohol andrecrystallised from water: M.P. (K) 177 -l8l C.

(b) Monocitrate of In prepared in solution in a ethyl acetate andrecrystallised from n-propanol: M.P. (K') 1 64-168 C. [a] =+72.5(concentration: 1% in methanol).

(c) Acetyl derivatives: 3a-pyrrolidinyl-2l-acetoxy-Sflpregnane-ZO-oneIak) made by acetylation in pyridine and recrystallisation from ethylacetate. M.P. (1K): 132- 133 C. (decomp.).

(d) Picrate of the acetyl derivative, C H H N small yellow tufts,recrystallised from ethanol: M.P. (K) 215-216 C.

(e) The 20,8, 21-diol, or 3a-pyrrolidinyl-Sfi-pregnane- 205,21-diolIas), obtained by reduction with sodium borohydride of the hydroxyaminoketone 1a and identical with the product of Example -3 made by theLeuckart reaction: M.P. (K) 254-256 C. (with ultimate decomposition)after recrystallisation from methanol.

(B) The compound ilaf, 3a-pyrrolidinyl-2l-hydroxy- SB-pregnane-ZO-one,is equally well prepared in good yield by following the proceduredescribed under Example l0, but using as starting materialhydrodesoxycorticosterone (Hg) and purifying the free amine as set forthunder (A) above.

The second method which can be employed for preparation ofamino-steroids of the general Formulae I is treating a mixture of aketone having the general Formula II and an amine, in an inert diluentwith hydrogen in the presence of suitable hydrogenating catalysts.

Amines of various kinds can be used, more especially ammonia, the loweraliphatic primary and secondary amines, saturated cyclic amines such aspyrrolidine, morpholine, piperidine, the N-alkyl piperazines, andaraliphatic amines such as benzylamine.

In general, the amine (ammonia is here considered to be an amine) asused in excess with respect to the 3-keto steroid being treated. Whenusing non-volatile amines, a slight excess, up to 10% is adequate. Whenusing volatile amines including ammonia, the excess can be rather more.

A catalyst which can suitably be used is, in the first place, palladium,preferably on a support, especially carbon black; for example 5% ofpalladium upon carbon black is quite suitable. Platinum can also beused, particularly in the form of the oxide. This too may be upon asupport, such as carbon black. Raney nickel may also be used.

The 3-keto steroid, amine, and catalyst are preferably placed in adiluent which is inert under the conditions of carrying out thehydrogenation. The usual solvents employed for catalytic hydrogenationscan be used. Lower aliphatic alcohols, which need not be completelyanhydrous, are especially suitable. However, other organic solvents, forexample ethyl acetate, ether, or dioxane, may be used. The quantity ofdiluent employed is not critical for successful working, and the choicethereof is dictated primarily by convenience. Generally from 5 to 50parts by weight of diluent are used per part of 3-keto steroid, but suchlimits are not mandatory.

The hydrogenation can usually be carried out successfully withoutapplying pressure, but it can nevertheless be carried out in equipmentdesigned for maintaining a super atmospheric pressure, preferably below10 kg./cm. of hydrogen. Usually the process is carried out attemperatures below the boiling point of the diluent, preferably from 0to 65 C. and most frequently at the temperature of the laboratory.Nevertheless in some cases it may be advantageous to cool in order towork below room temperature, and in other cases, to heat. As a rule thehydrogenation can be continued until hydrogen ceases to be absorbed oruntil the calculated amount has been absorbed, taking into account incertain cases what may be required to saturate any double bonds, or, ifit be desired, to effect or avoid hydrogenolysis (for example, to removethe benzyl group in the case of benzylamine). Taking into account theserequirements the reaction time may vary from several hours to severaldays.

Under the mild hydrogenation conditions envisaged above one can largelyor wholly -avoid the reduction of carbonyl groups, other than that atposition 3, which may be present in the 3-keto-steroid employed asstarting material.

In many cases only the 35 isomer can be isolated from the hydrogenationproduct by crystallisation to the exclusion of any other isomer.

When the amine used is benzylamine, it has been found that if thehydrogenation is'not stopped when the requisite amount of hydrogenrequired to produce the 3-benzylamine compound has been absorbed, theabsorption continues, particularly when working above room temperature.Hydrogenolysis, accompanied by fission of the benzylamino group andformation of the corresponding primary amine, then occurs.

The invention includes such mode of carrying it into effect, that is tosay, in the case of benzylamine, hydrogenation carried to the stage ofhydrogenolysis. This enables the ready production of the correspondingprimary amines, especially those having a primary amino group having the,8 configuration.

Certain primary amines are difiicult to isolate as the free base,perhaps by reason of a condensation between the NH group and a CO group,so long as another is present in the molecule.

The applicants have found that it is particularly convenient, in orderto isolate the primary amine in a pure state and in good yield, to causeit to react with an appropriate aldehyde in order to convert it into aSchifis base, so long as it is easy subsequently to regenerate theprimary amine from that base. Benzaldehyde is very convenient for thispurpose. Regeneration of the primary amine can be effected either byhydrogenating the Schiffs base, or by hydrolysis with a mineral acid.This method of producing and purifying the primary amine is, in fact, avaluable means of preparing it, although it can also be prepareddirectly, using ammonia and hydrogen, although the difliculties ofisolating and purifying it are then greater. These difiiculties are lessin evidence when in the case of unsaturated 3-keto steroids, such asprogesterone, the starting materials are the corresponding reducedcompounds, such as pregnanedione.

The following examples illustrate the second procedure for synthesis ofaminated steroids of type I (catalytic reductive amination). Thereferences such as Ia, IIa, refer to the formulae and tables givenabove.

EXAMPLE 13 3B- and 3a-pyrrolidinyl-5a-pregnane-ZO-ones (Ia and Id) Thereare charged into an apparatus for catalytic hydrogenation at ordinarypressure, fitted with an electromagnetic stirrer:

3.15 g. (0.01 mol) of 5a-pregnane-3,20-dione (IIb) M.P. 1171l9 C.), 0.9ml. of pyrrolidine (0.011 ml.),

105 ml. of absolute alcohol and 0.3 g. of palladium supported oncharcoal and containing palladium. The mixture is hydrogenated at roomtemperature. After about an hour the theoretical amount of hydrogen (224ml.) is taken up. The catalyst is then filtered off, the filtrateevaporated to dryness, the residue taken up in petroleum ether andallowed to crystallise. 1.5 g. of a white product, M.P. 120125 C. (tube)is obtained which is recrystallised from petroleum ether. It is3fl-pyrrolidinyl-5a-pregnane-20-one (Ic) M.P. 127129 C. (tube)(concentration=1% in chloroform). The melting point is not depressedwhen mixed with the base made by the formic acid reduction of thepyrrolidinyl enamine prepared from 5a-pregnane-3, 20-dione (see Example11). It has an infrared spectrum which is identical with that of thislatter base (determined in solution in carbon disulphide) When added toan aqueous solution of furmaric acid, the base gives a hydrogenfurmarate, which is recrystallised from water: white crystals, M.P.72-75 C. (tube).

The petroleum ether mother liquors remaining after isolating the3,9-amine are evaporated to dryness. The residue is dissolved in alcoholand 1 g. of furmaric acid is added to the solution. The fumarate formedis precipitated by adding ether plus petroleum ether. It is filtered offand recrystallised, firstly from isopropanol and then from water, giving0.3 g. of the hydrogen fumarate C H ON, C H O ,M.P. (tube) 209212 C.

The corresponding base is liberated from this fumarate by means ofammonia. After extracting with ether, washing the ether solutions with,drying them over sodium sulphate, filtering and evaporating the solvent,the regenerated amine is recrystallised from methanol. It is3a-pyrrOIidinyI-Sa-pregnane-20-one (Id), M.P. (tube): 102-104 C. [a]=+82.6 (concentration 0.5% in chloroform).

EXAMPLE 14 35- and 3a-pyrrolidinyl-Sfl-pregnane-ZO-ones (Ia and lb) (a)Starting from 5/3-pregnane-3,20 dione (He):

The process is carried out exactly as with the sd-H derivative, using3.15 g. (0.01 mol.) of 5fi-pregnane-3,20- dione (110), but instead ofevaporating to dryness after filtering off the catalyst, the filtrate isconcentrated to 25 cm. and allowed to crystallise. On cooling there isobtained a first crop of 3fi-pyrrolidinyl-Sfl-pregnane-20-one (Ia),white crystals, M.P. (tube) 132-133 C. (yield 1.35 g.). After filteringconcentration of the mother liquors affords 0.15 g. of the same product,M.P. (tube) 131-132 C. [a] =+98.1 (concentration 1% in chloroform)Addition of fumaric acid to a solution of this base in anhydrous acetonegives a hydrogen fumarate. The latter is recrystallised from anhydrousacetone M.P. 198-201 C. (tube).

The alcoholic mother liquors after isolating the 3,8-pyrrolidinyl-Sfi-pregnane-ZO-one are treated with fumaric acid to obtainthe salt. The fumarate is precipitated by a mixture of ether and petrolether. The crystals obtained are filtered off and recrystallised fromacetone and then from water, affording 0.7 g. of fumerate (M.P. (tube)172-174 C.). The base is liberated from the latter by using ammonia, asdescribed in Example 13 and the regenerated amine recrystallised frompetrol ether. This gives 0.3 g. of a compound (M.P. 116-118 C. (tube))which shows an unchanged mixed melting point with3a-pyrrolidinyl-5fi-pregn-ane-3,20-dione, Ib, made as described inExample 10. The infrared spectra, determined in carbon disulphide, aresuperimposable.

(b) Starting from progesterone (11a):

The following are charged into a hydrogenation apparatus for use atatmospheric pressure which is fitted with an ele tromag etic stirrer=5.2 g. of progesterone (Ila), ml. of absolute alcohol, 1.5 cm?pyrrolidine and 0.5 g. of 5% palladium supported upon charcoal. Thetheoretical amount of hydrogen is absorbed in 20 minutes at roomtemperature.

The product is filtered, the filtrate evaporated to drymess, the residueis dissolved in 20 ml. of ethanol and allowed to crystallise slowly. Inthis way there is obtained 2.65 g. of a white product (M.P. 128-133 C.(tube)), which after recrystallisation from ethanol, melts at 132- 133C. (tube) and shows no depression of melting point with3fi-pyrrolidinyl-SB-pregnane-20-one, made by process (a). Furthermorethe infrared spectra as determined in carbon disulphide, aresuperimposable.

EXAMPLE 15 3,B-dimethylamine-Sfl-pregnane-ZO-one (If) The reactionmixture consists of: 0.00 33 mol (1.05 g.) of 5B-pregnane-3,20-dione(He), 30 ml. of absolute alcohol, 5 ml. of a 20% alcoholic solution ofdimethylamine, and 0.1 g. of 5% palladium supported upon charcoal.Hydrogenation is carried out at ordinary pressure and 40 -45 C. using anelectromagnetic heating stirrer. Reaction takes place very slowly (5 to6 hours). When it is finished (after 74 cm. of hydrogen have beenabsorbed) the catalyst is removed by filtration, the filtrate evaporatedto dryness, the residue taken up in absolute alcohol and 0.4 g. offumaric acid added. The fumarate formed is filtered off andrecrystallised from alcohol; yield 0.6 g. It is the acid fumarate of3/3dirnethylamino-5fipregnane-ZO-one, M.P. (tube) 212-214 C.

(concentration 1% in methanol).

The free base is obtained from the hydrogen fumarate in the usual wayand melts at 9697 C. (tube). Examination of the infrared spectrum ofthis amine as determined in carbon disulphide solution, shows thepresence of bands characteristic of ZO-ketones, with a normal Cconfiguration (R. Norman Jones and coworkers. J. Amer. Chem. Soc., 1955,volume 77, page 651) with an axially (or 13) disposed tertiary aminogroup at position 3 (triplet in the region 7.2-7.6,u. and absorption at11.60 with a minimum at 11.40;.t).

EXAMPLE 16 3 3-piperidinyl-5/3-pregnane-20-one (Ig) The reaction mixturecomprises: 5fi-pregnane-3,20- dione (IIc; 0.02 mol), dry piperidine (2.4ml., 0.022 mol), 5% palladium supported upon charcoal (0.1 g.) andabsolute alcohol (50 ml.). This is hydrogenated at ordinary pressure and50 C. making use of an electromagnetic stirrer which heats. The reactionis very slow: it requires more than 24 hours for completion. Afterfiltration and concentration, the 3B-piperidinyl-5fl-pregriane-ZO-one(Ig) crystallises; yield=3.60 g., M.P. (tube): 116-119 C. Afterrecrystallisation from absolute alcohol it melts at 121l22 C. (tube).[a] =+94.2 '(concentration=1% in chloroform).

EXAMPLE 17 3B- and 3a-morpholinyl-SB-pregnane-ZO-one (Ill and Ii)Hydrogenation is carried out exactly as in Example 16, but thepiperidine is replaced by 1.9 g. (0.022 mol) of dry morpholine.Absorption is even slower, After 34 hours, only 328 cm. of the 448 cm.of hydrogen theoretically required have been absorbed. So afterfiltering and evaporating the filtrate to dryness in vacuo, the residualoil is itself dissolved in 5 ml. of acetic acid and 250 ml. of water arethen added. The amines remain in solution as salts, and the non-basicproducts (2.4 g.) can be extracted with ether. The aqueous solution ofamine acetates is then made alkaline and the free bases are extractedwith ether. The ethereal extract is washed with water and dried oversodium sulphate- After filtering and evaporating to dryness, thecolourless oil obtained (yield 5.2 g.) is taken up in petroleum etherand left to crystallise slowly to give 3.4 g. of a white product M.P.(K) 142-144 C. which is recrystallised from petroleum ether. It is3fi-morpholinyl-SB-pregnane-ZO-one (Ih), M.P. (tube)=l44-145 C. [a].=+89.7 (concentration=1% in chloroform).

The petrol ether mother liquors are taken down to dryness and theresidue thus obtained (1.6 g.) is made acid with a solution of 0.5 g.fumaric acid dissolved in 4.5 cm. of isopropanol. This affords agelatinous fumarate, which is repeatedly crystallised from acetone.Finally there is obtained 1.0 g. of white crystals, M.P. (tube) 164166C. (decornp) This fumarate is decomposed with ammonia and the free base,worked up in the usual way, is recrystallised from light petroleumether. It is 3a-morpholinyl-SIB-pregnane-ZO-one '(Ii), M.P. (tube)136-137, [u] =+103 (concentration=1% in chloroform).

EXAMPLE 18 3pand 3a-(4-methyl-1-piperazinyl)-5B-pregnane-20-ones (Ij and1k) These amines are prepared in the same way as the correspondingmorpholine derivatives, but using 2.2 g. of N-methylpiperazine in placeof the morpholine. In this case also the absorption of hydrogen is veryslow: even after 54 hours the theoretical amount of hydrogen has notbeen taken up. The basic fraction obtained yields, in petroleum ether,3.1 g. of white crystals, having a double melting point: 159160 C. and166167 C, (tube). It is 3(3-(4-methyl-1-piperazinyl)-5fi-pregnane-20-one(Ij). The specimen of constant melting point is recrystallised frompetroleum ether; double melting point (tube) 162- 164 C. and 168-169" C.[a] =+9O.6 (1% concentration in chloroform).

The mother liquors are taken down to dryness, and the residue, aftersolution in acetone, is converted into a salt by addition of a methanolsolution of fumaric acid. The fumarate which precipitates isrecrystallised from 95% alcohol. The base is liberated from thisfumarate (1.70 g., M.P. (K) 270 C.) with ammonia, and it is then workedup in the usual way. It is 3a-(4-methyl-1-piperazinyl)-5fi-pregnane-20-one (1h), M.P. (tube) 147- 149 C. (fromlight petroleum ether); [a] =+83.5 (concentration: 1% in chloroform).

EXAMPLE 19 3/3-methylamine-5fl-pregnane-20-one (ll) 6.3 g. (0.02 mole)of 5,8-pregnane-3,20-dione (He) is dissolved in 120 ml. of absolutealcohol. Then 30 ml. of a 20% by weight solution of methylamine inabsolute alcohol are added, together with 0.1 g. of 5% palladiumsupported upon charcoal. The theoretical amount of hydrogen is absorbedafter 8 /2 hours, working at 50 C. and using an electromagnetic heatingstirrer. The mixture is then filtered, the filtrate evaporated todryness and the residue recrystallised from di-isopropyl ether and thenfrom methanol. The crystals thus obtained do not have a sharp meltingpoint, and they are then distilled from an oil bath; B.P. at 0.05 mm. ofmercury 215 C. (bath temperature). It is3B-methylamino-5[3-pregnane-20-one (ll) M.P. (tube) 8182 C.

EXAMPLE 20 3 fi-benzylamino-5fl-.pregnane-20-one (Im) (a) Using apalladium catalyst: The following mixture is introduced into apparatusfitted with an electromagnetic stirrer for carrying out hydrogenation atordinary pressure; 25.2 g. of progesterone (0.08 mol), 9.6 gm. ofbenzylamine (0.088 mol), 200 ml. of absolute alcohol and 2.4 g. of 5%palladium supported upon charcoal. After 9 hours at room temperature3,900 cm. hydrogen is absorbed.

A white product crystallises out and is deposited on the catalyst. Themixture is filtered and the mass of catalyst is extracted withchloroform or hot benzene. On evaporation of the solvent3B-benzylamin0-5fi-pregnane 20-one crystallises out (yield 21 g.: yield64.3%). After a single recrystallisation from ethanol the M.P. isconstant at 138140 C. (tube), yield 16.1 g. (very nearly 50%), lD =+85.1(concentration 1% in chloroform). The infrared spectrum (determined incarbon disulphide solution) shows the normal 20-keto group of the seriesand a monosubstituted aromatic ring.

(b) Using a platinum oxide catalyst: 6.3 g. (0.02 mol) of progesterone,0.63 g. of platinum oxide (Baker Ltd.), 1.2 g. of purified [animalcharcoal, 2.45 ml. of benzylamine and 50 ml. of absolute alcohol arehydrogenated at room temperature using electromagnetic stirring. Whentwo equivalents of hydrogen have been absorbed (in 4 /2 hours), aproduct crystallises out from the reaction mixture, but absorptioncontinues at an appreciable rate. The reaction is stopped after 22 hoursand the reaction mixture filtered. Then, after adding ammonia so thatcolloidal platinum does not pass into the filtrate, the steroid retainedby the filter together with the catalyst, is extracted with chloroform.The chloroform solutions are then evaporated in vacuo to dryness (4.83g.). The dry residue is recrystallised from absolute alcohol. Thematerial thus obtained melts at 136-139 C. (K) and does not show anydepression of melting point when admixed with3fi-benzylamino-SB-pregnane-ZO-one obtained as described under (a)above. Furthermore the infrared spectra are substantiallysuperimposable, although certain differences in the region from 8 to9 1. lead one to suspect that isomers with a S-a-H configuration arepresent in appreciable amounts.

(c) With Raney nickel catalyst: A mixture of progesterone (6.3 g.; 0.02mol) freshly-prepared Raney nickel (3 g.), benzylamine (2.45 ml.) and 50ml. of absolute alcohol is hydrogenated in a shaking apparatus.Absorption of gas is rather slow: about 15 hours are needed for thesteroid to take up two equivalents of hydrogen. The latter crystallisesout in the reaction mixture. The steroid precipitated along with thecatalyst is extracted with chloroform. After evaporation in vacuo, theresidue is dried and then weighs 5.1 g. It is recrystallised fromabsolute alcohol: M.P. (K) 136-138 C. shows no depres sion of meltingpoint on admixture with the product prepared according to (a).

From the mother liquors 0.26 g. of a product M.P. (K) 250 C. can berecovered.

EXAMPLE 21 3paamino-SB-pregnane-ZO-one (lo) and derivatives thereof (a)Via the derivative Is containing a benzylidene group:

Into an apparatus for hydrogenation at ordinary pressure, fitted with anelectromagnetic stirrer which heats, there is introduced 6.3 g. of5B-pregnane-3,20-dione (0.02 mol, He), ml. of absolute alcohol, 2.4 g.(0.022 mol) of redistilled benzylamine and 0.6 g. of 5% palladiumsupported upon charcoal. Throughout the entire hydrogenation (5 hours)the mixture is heated to 50 C. and slightly more than two equivalents ofhydrogen (1,020 cm. compared with the theoretical quantity of 896 cm?)are absorbed. The mixture is filtered and 2.4 g. (0.022 mol.), ofbenzaldehyde is added to the filtrate. The whole is then concentrated atraised temperature until crystallisation begins: this gives 5.4 g. ofthe benzylidene derivative Is. M.P. (tube) 188190 C.

A specimen of the compound Is, 3 8-benzylidene amino- Sfipregnane-ZO-one, is recrystallised from ethyl acetate M.P. (tube) 197198C.

The crude 'benzylidene derivative, Is, is catalytically hydrogenatedunder conditions identical with those described above, in the presenceof 5% palladium supported upon charcoal and ml. of absolute ethanol.After 2.5 hours it has absorbed substantially the quantity of hydrogenrequired to split the regenerated benzylamine derivative. The product isfiltered, the filtrate evaporated to dryness in vacuo, and the residuedissolved in boiling petroleum ether and then left to crystalliseslowly. In this way there is obtained 2.0 g. of large colourlesscrystals, M.P. (tube) 120l22 C. while from the mother liquor there isrecovered a further 1.2 g. of product. M.P. (tube) 95105 C., butundergoing change when attempt is made to purify it byrecrystallisation. The product M.P. 120122 C. has [a] =+98.4-(concentration: 1% in chloroform). The infrared spectrum, determined inpetrolatum has little or no absorption in the region 6.25 to 6.30 4,unlike 3a-amino steroids. But it has a very sharp absorption in theregion of 14.70u.

The N-acetyl derivative of this amine is obtained in quantitative yieldby the action of acetic anhydride on the free base. This affords whitecrystals, Ir which after recrystallisation from di-isopropyl ether hasM.P. (tube) 127-128 C. [u] =+9O.7 (concentration 1% in ethanol).

(b) Via the acid fumarate:

To 5.15 g. of pregnanedione IIc are added 165 ml. of absolute alcohol, 3g. of benzylamine and 0.5 g. of 5% palladium supported upon charcoal.Hydrogenation is carried out at 40-43" C. and continued until an amountof gas sufficient to effect hydrogenolysis of the intermediatebenzylamine derivative Im, has been absorbed. The product is filteredand the primary amine separated as the acid fumarate, yield 2.4 g. M.P.(tube): 176- 178 C. It is very difficult to purify. By the action ofammonia upon this fumarate the free base is obtained and is extractedwith cyclohexane. The cyclohexane solutions are washed with water anddried over sodium sulphate. Evaporation yields the amino steroid, whichis repeatedly recrystallised from petroleum ether. This purification isaccompanied by copious resinification and large losses. It has not beenpossible to raise the melting point of the 3 8-amino-Sfl-pregnene-ZO-one(I) above 112-114 C. (tube).

(0) By hydrogenolysis of the purified benzylamino derivative Im, thelatter being prepared from progesterone (see above) 16.1 g. of purified3-benzylamino-5B- pregnane-20-one, M.P. 138-140 C. (tube) made byreductive catalytic amination of 25.2 g. of progesterone in the presenceof palladium (see above), are suspended in 200 ml. of absolute alcoholand stirred electromagnetically in an atmosphere of hydrogen at ordinarypressure and 50 C. in the presence of 1.6 g. of palladium supported oncharcoal. Hydrogenolysis is practically complete after 4 hours, when thetheoretical amount of hydrogen (900 m1.) has been absorbed. The catalystis removed by filtration and the alcohol removed in vacuo at 50 C. Theresidue is dissolved in 40 ml. of boiling petroleum ether. The solutionis allowed to crystallise slowly to give, after drying, 11.0 g. of3p-amino-5i3- pregnane-ZO-one, I0. M.P. 118-120 C. (tube). The meltingpoint is not depressed when admixed with the product obtained by method(a). The yield is 74% (or 37% based on the progesterone).

In an analogous experiment the amine was isolated as the acid maleate,by adding a concentrated ethanol solution of maleic acid to the filteredcrude hydrogenolysis product. Recrystallisation from water gave theanalytically pure salt. M.P. (tube) 184-186 C., [a] =+7l.9(concentration=1% in 96% alcohol).

(d) Preparation of the ascorbate of 3fi-amino-5fipregnane-20-0ne, Io byhydrogenolysis of the purified benzylamine derivative (Im), the latterbeing made from progesterone (see above).

20.0 g. of purified 3/3-benzylamino-Sfi-pregnane-ZO-one (Im). M.P.138140 C. (tube), are suspended in 200 ml. of isopropanol and stirredelectromagnetically at 50 C. in hydrogen at ordinary pressure, and inthe presence of 2 g. of 5% palladium supported upon charcoal.Hydrogenolysis is practically complete after 5 hours (1120 ml. hydrogenis absorbed). The catalyst is removed by filtration. The filtrate isheated to boiling, under an atmosphere of nitrogen and a boilingsolution of ascorbic acid (10 g.) in methanol (75 ml.) added.

The methanol is evaporated and the ascorbate" crystallises. Aftercooling, the white crystals are filtered off and sucked dry. They aredried in a high vacuum. The yield obtained 20.8 g., is equal to 86%based on the benzylamine derivative, Im, and 43% based on theprogesterone. M.P. (tube) 179181 C.,

(concentration: 0.5% in water).

This ascorbate is one of the few salts of base I0 which is readilysoluble in water.

(e) By attempted hydrogenolysis of the anisylidene derivative (Iv).Isolation of SB-(methoxy-phenylmethylamino)-5;8-pregnane-20-0ne (Ix).

A mixture of 5B-pregnane-3,20-dione (II c; 6.3 g.) and benzylamine (2.4g.) is hydrogenated as described above under (a). The crude primaryamine formed by hydrogenolysis is condensed with anisaldehyde (4.4 g),by heating for 1 hour under reflux in the presence of alcohol. Afterconcentrating to ml., 6.3 g. of Schifis base containing an anisylidenegroup (Iv) is isolated by crystallisation. M.P. (tube): -177 C.Analytically pure imine is obtained by recrystallisation from ethylacetate, M.P. 179180 C. (tube).

2.2 g. of this derivative (Iv) is treated, in absolute alcohol at 50-52C. with hydrogen at ordinary pressure, in the presence of 5% ofpalladium supported upon charcoal (0.2 g.). Actually a single equivalentof hydrogen is absorbed (135 ml., whereas 226 ml. would be required forhydrogenolysis). After filtering off the catalyst, evaporation todryness in yacuo and crystallisa tion from boiling petroleum ether, thesecondary amine Ix is obtained M.P. (tube) l24-125 C.

(concentration 1% in ethanol).

EXAMPLE 22 35- and 3a-aminO-Sa-pregnane-20-ones (lq and 1A) andderivativesIsolation of funtumine (a) Using benzylamine as a source ofnitrogen;

The following are charged into a hydrogenation apparatus, fitted with anelectromagnetic stirrer which heats:

6.3 g. of 5a-pregnane-3,20-dione (IIb; 0.02 mol), 80 ml. of absolutealcohol, 2.4 g. of benzylamine (0.022 mol), and 0.6 g. of 5% palladiumsupported upon charcoal. The mixture is stirred for 5 hours at 50 C.after which 960 ml. of hydrogen have been absorbed (896 ml. istheoretically required to bring about reduction and hydrogenolysis). Thecatalyst is filtered off, 2.4 g. of benzaldehyde (0.022 mol) is added tothe filtrate, the mixture is concentrated to half its volume and thenallowed to crystallise. In this way there are obtained 3.5 g. of 35-benzylidene-amino-5a-pregnane-20-one (It) M.P. (tube) 197 C.Recrystallised from ethyl acetate it weighed 2.5 g. and had M.P. 202-204C. (tube).

The benzylidene derivative (It) purified in this way is hydrogenated atordinary pressure and 50 C. in 80 ml. of alcohol containing 0.2 g. 5% ofpalladium supported upon charcoal. In two hours the amount of hydrogentheoretically required to form the free amino group is absorbed. Afterfiltering off the catalyst, the alcohol is evaporated in vacuo. Theresidue crystallises. By dissolving it in boiling cyclohexane and slowlyevaporating pure 3,B-amino-5ot-pregnane-20-one (Iq) is obtained, M.P.(tube) 157158 C. weight 1.45 g., [a] -=+96.3 (concentration=1% inchloroform). The infrared spectrum, as determined in petrolatum exhibitsbands characteristic of equatorial compounds.

From this base there was prepared an acid fumarate, M.P. (tube). 232-234C. after recrystallisation from methanol; and an imine Iw by reactionwith anisaldehyde (prepared in alcohol and recrystallised from ethylacetate) M.P. (tube) 193 194 C.

The several mother liquors obtained after separation of the benzylidenecompound (II), M.P. 202-204 C. (and belonging to the 3B-amino series)are evaporated to dryness in vacuo. The residue obtained is dissolved indiisopropyl ether or di-ethyl ether and the solution treated overnightwith dilute hydrochloric acid. In this way the imine is hydrolysed and asparingly soluble amine hydrochloride is precipitated. It is purified byrecrystallisation from a chloroform-acetone mixture. It is funtuminehydrochloride, or the hydrochloride of 3a-amino-5a-pregnane- 20-one.M.P. (micr.) 260-265 C. yield 1.4 g.

This hydrochloride or treatment with ammonia affords the base funtumine(IA) which dissolves in the ether. After decanting the separatedethereal extracts are washed with water and then dried over sodiumsulphate. The ether is then removed in vacuo and the residual oilobtained crystallised from a mixture of ether and light petroleum (B.P.3545 C.). In this way 1.1 g. of the base, M.P. (tube) 113 1l7" C. isobtained.

The pure specimen for analysis of funtumine is obtained by anotherrecrystallisation from ether-light petroleum ether. Its constants arepractically identical with those of the product isolated by Ianot andcoworkers from Funtumia latifolia (Apocyanacae). It is3a-amino-5u-pregnane-20-one (funtumine, IA) M.P. (tube) 120-124 C. [a]=+l03.l (concentration=l% in chloroform). The infrared spectrum asdetermined in petrolatum confirms this configuration.

By treating funtumine in alcoholic solution with benzaldehyde (30minutes refluxing) an imine containing a benzylidene group is formed. Itis crystallised from petroleum ether and then from methanol. It isBet-benzylidene imino-Sa-pregnane-ZO-one (Iu), [a] =+96.9 (concentration1% in CHCl M.P. 200-202 C. (tube). It gives a depressed mixed meltingpoint with the isomeric 3fl-benzy1idene imino compound (It) describedabove.

(b) With ammonia, NH as a source of nitrogen:

An apparatus for hydrogenation at ordinary pressure fitted withelectromagnetic stirrer which heats is charged with 6.3 g. of5a-pregnane-3,20-dione (IIb) 0.02 mol, dissolved in 100 ml. of absolutealcohol, 20 ml. of a 7% solution of gaseous ammonia in absolute alcohol,and 0.6 g. of 5% palladium supported upon charcoal. Absorption ofhydrogen is slow: 11 hours are required for absorption of 480 ml. ofgas, this quantity being slightly more than the theoretical amount (448ml.). The catalyst is separated by filtration. It retains crystals of anorganic product which are dissolved in chloroform, but there is no aminein the residue obtained on evaporating the chloroform. The bases formedin the reaction remain in solution in alcohol. This solution onevaporating to dryness in vacuo, gives a residue weighing 5.3 g., whichis dissolved in the smallest amount of alcohol and then converted to aSchifis base by reaction with benzaldehyde. The benzylidene derivative,which separates on rubbing the walls of the vessel, is isolated: weight1.9 g., M.P. (K) 190192 C. It is recrystallised from ethyl acetate. The3;.8-benzylamino-5a-pregnane-20-one thus prepared (It) has M.P. (tube)19820l C. It does not lower the M.P. 202-- 204 C. (tube) of fusedproduct obtained as described under (a) above.

EXAMPLE 23 3p-pyrrolidinyl-5a-androstane-175-01 (Iz) 5.8 g. ofandrostanolone (IId: .020 mol) and 1.8 ml. of pyrrolidine (0.022 mol)dissolved in 210 ml. of absolute ethanol are hydrogenated at ordinarytemperature and pressure, in the presence of 0.6 g. of 5% palladiumsupported upon charcoal. The theoretical amount of hydrogen (450 ml.) isslowly absorbed. The catalyst is filtered off, the filtrate concentratedto 75 ml. and left to cool slowly.3,8-pyrrolidinyl-5a-androstane-17,8-01 (Iz) 26 crystallises out, yield4.3 g., M.P. l82-184 C. (tube). It is obtained analytically pure byrecrystallisation from ethyl acetate, M.P. (tube) l79l82 C., [a] =+8.8(concentration: 1% in chloroform). The steric structure is settled byanalogy.

EXAMPLE 24 3fidimethylamino-5u-androstane-l7 3-ol (lab) This is preparedin the same way as the preceding derivative (Iz). Upon replacing thepyrolidine by 30 ml. of a 20% alcoholic solution of dimethylamine.

A product of constant melting point (yield 2.2 g.) is obtained after twocrystallisations from ethyl acetate M.P. (tube): 172174 C., [a] =+8.8(concentration: 1% in chloroform).

The methiodide of compound Iab is made by dissolving the base (1.5 g.)in absolute alcohol (15 ml.) and then adding an excess methyl iodide.The quaternary salt crys tallises out spontaneously. The white crystals,which turn yellow in air, are filtered off, sucked dry andrecrystallised from ethyl acetate. Yield 0.5 g. M.P. (K): 234- EXAMPLE25 3,8-pyrrolidinyl-Sfi-androstane-175-01 (Iaa) 2.9 g. of testosterone(0.01 mol) and 0.9 ml. of pyrrolidine (0.011 mol), dissolved in 50 ml.of absolute alcohol, are hydrogenated =at ordinary temperature andpressure, using 0.15 m. of a 5% palladium catalyst supported uponcharcoal.

After absorption of the theoretical amount of hydrogen (448 ml), themixture is filtered and concentrated. The3B-pyrrolidinyl-Sfiandrostane-1718-01 formed is recrystalflised fromethyl acetate. Yitld 0.5 g. M.P. (K): 234- 236 C., [a] =-+23.5(concentration: 1% in chloroform).

EXAMPLE 26 21-hydroxy-3,8-pyrro1idinyl-5 fi-pregnane-20-one (Iae) Thefollowing are charged to a hydrogenation flask: 5.5 g. ofhydrodesoxycorticosterone (IIg), ml. of absoute alcohol 1.5 ml. ofpyrrolidine and 0.3 g. of 5% palladium supported upon charcoal. Themixture is stirred electromagnetically at ordinary temperature andpressure. After the theoretical amount of hydrogen (373 ml.) has beenabsorbed, the catalyst is filtered off and the filtrate evaporated todryness in vacuo.

The crude 21-hydroxy-3B-pyrrolidinyl-S S-pregnane-ZO- one (Iae) yield3.5 g., separates as white needles when recrystallised from diisopropylether M.P. 149-151 C. (yield 2.2 g.). An analytically pure specimenmelts at 152154 C. (tube) [u] =+86.2 (concentration: 1% in chloroform).

It depresses the melting point of the 3oc-iSOII16I, which is obtained bythe formic acid reduction of the pyrrolidine enamine ofhydrodisoxycorticosterone (see Example 12) which has M.P. 157159 C.(Iaf).

EXAMPLE 27 3fi-pyrrolidinyl-5/8-pregnane-20fi,21-diol (Iar) Theprocedure of Example 26 is followed, but instead ofhydrodesoxycorticosterone, 205,2l-dihydroxy-SB-pregnane-3-one (IIh) isused.

Recrystallisation from diisopropyl ether gives 1.8 g. of3fi-pyrrolidinyl-5fi-pregnane-20,t3,2l-diol (Iar) M.P. (K) 188l90 C.,[a] =+l7.5 (concentration: 0.7% in chloroform) EXAMPLE 28 2l-acetoxy-38-amino-SB-pregnane-ZO-one (Iam) derivaderivatives (a) Fromdesoxycorticosterone acetate (Hi) the following mixture is hydrogenatedat ordinary pressure: 22.2 g. (0.06 mol) of desoxycorticosterone acetate(Hi), 320 ml. of absolute alcohol, 7.2 g. of 'benzylamine (0.06 mol) and2.2 g. of palladium supported upon charcoal.

After about 7 hours 0.12 mol (2830 ml.) of hydrogen is absorbed ineffecting saturation of the doublebond and reductive amination. Themixture is then heated to 50- 55 C. The absorption, which had slackenedoff very much, starts up again (hydrogenolysis of the intermediatebenzylamino compound), and 0.06 mol of hydrogen is found to be absorbedin 5 hours. The catalyst is filtered off, 8 g. of benzaldehyde is addedto the filtrate and the mixture is then concentrated to half its volume.The imine formed (Inc) is allowed to crystallise out. It is filtered offand recrystallised from absolute alcohol. Yield 15.0 g. M.P. (tube):l28l31 C.

A further recrystallisation gives2l-acetoxy-3fl-benzylideneamino-Sfi-pregnane-ZO-one (Iao) of constantmelting point. The melting point is 132134 C. (tube).

(concentration: 1% in chloroform).

4.6 g. of the benzylidene derivative (Iao) M.P. (tube): l28-131 C. ishydrogenated at ordinary pressure and 50 C. in the presence of 80 ml. ofabsolute alcohol and 0.5 g. of 5% palladium supported upon charcoal.

After the theoretical amount (448 ml.) of hydrogen has been absorbed (2hours), the catalyst is filtered off, the alcohol removed byevaporation, and the residue taken up in ether. The latter is partiallyevaporated whereupon 0.9 g. of 2l-acetoxy-3fi-amino-5ti-pregnane-20-one(Iam) M.P. (tube) 163 167 C. is obtained.

Purification by recrystallisation from ether or ethyl acetate is adelicate operation, since the product undergoes change. M.P. (tube)l76-178 C., [u] =--}-100.4 concentration=l% in chloroform).

The N-acetyl derivative of this amine is made by the action of aceticanhydride on the free base: the yield is quantitative. Recrystallisationfrom diisopropyl ether gives white crystals (Ian), M.P. (tube) 129131 C.

(concentration: 1 in chloroform).

Better yields of the free amine Iam are obtained by precipitating it asthe acid maleate after filtering oflf the catalyst. This is done byadding a solution of maleic acid in ethyl acetate. There is thusobtained 2.7 g. of the crystalline maleate (M.P. (tube) 171-173 C.)which is purified by recrystallisation from cold water, M.P. (tube)178-l80 C. [a] ==+80.2 (concentration 1% in chloroform) (b) Fromhydrodesoxycorticosterone acetate (IIj).

The method used above is followed using 3.75 g. of Hi (0.01 mol)dissolved in 80 ml. of hot absolute alcohol, 1.2 g. of benzylamine and0.44 g. of 5% palladium supported upon charcoal. After absorption of0.02 mol of hydrogen, removal of the catalyst by filtration and additionof 1.5 g. of benzaldehyde, there is obtained 3.8 g. of the benzylidenederivative Iao M.P. (tube) 125 'l27 C. identical with the product madeby procedure (a).

EXAMPLE 29 Reductive arnination, using benzylamine, of 20fl,2l-dihydroxy-5/3-pregnane-30-one (Ilh) 1.5 g. of compound II]: (M.P.(K) 160-162 C.) is dissolved in 25 ml. of absolute alcohol, and 0.55 ml.benzylamine plus 0.15 gm. of 5% palladium supported upon charcoal.Hydrogenation is effected at 50 C. under ordinary pressure, using anelectromagnetic stirrer which heats. Absorption of hydrogen is very slowand after 30 hours, only 115 ml. of hydrogen have been absorbed. To addthe amount theoretically required for reductive amination and subsequenthydrogenolysis (200 ml.), fresh catalyst (0.15 g.) must be added andstirring continued at 50 for a further 12 hours. The mixture isfiltered, the residue washed with boiling alcohol and the filtrateevaporated in vacuo. The residual'oil is crystallised from ether 28weight 0.67 g. M.P. (K) l32136 C. Attempts at recrystallisation failedas the product was very unstable. This crude-BB-amino-Sfl-pregnane-Z05-, .2l diol (Iax) is dissolved in alcohol andheated for 1 hour on a boiling water bath with a slight excess ofbenzaldehyde. The solution obtained is concentrated: then, on rubbingthe walls of the vessel, the benzylidene imine (Iay) crystallises. It ispurified by solution in hot alcohol and then has M.P. (K) l56-158 C. Thesubstance thus obtained is still not analytically pure.

EXAMPLE 30 3-pyrrolidinyl-l7u,21-dihydroxy-5-pregnane-l 1,20- dione (Iaiand Iaj) 4.8 g. cortisone (IIk, 0.0132 mol) is dissolved in 140 ml. ofabsolute alcohol and 1.2 ml. of pyrrolidine and 0.30 g. of 5% palladiumsupported upon charcoal added. Hydrogenation is effected at ordinarytemperature and pressure, 630 ml. hydrogen being absorbed (theoretical584 ml.). The mixture isfiltered and the filtrate concentrated in vacuoat 35 C. using a capillary tube bubbler connected to a source of drynitrogen. The residue is dissolved in dry ether under an atmosphere ofnitrogen. In this way 1.65 g. of a rather unstable crystalline productis obtained, M.P. (K) 208 C. It is recrystallised with difficulty fromabsolute ethanol: it is 3-pyrrolidinyl-17a,21-dihydroxy-Sg-pregnane-l1,204iione (Iai), in the form of its hemihydrate,M.P. (tube) 228230 C. [a] +74.7 (concentration 1% in chloroform). It isbelieved that the two .5 orientations are Zia-amino and 55H.

The ether mother liquors obtained after isolating the above hemihydrate,are evaporated to dryness in vacuo, and the residue taken up in absolutealcohol. Treatment with concentrated hydrochloric acid gives thehydrochlorides of the amines present. The mixture is filtered and thecrude hydrochloride triturated with absolute alcohol. This affords 3 g.of salt sparingly soluble in ethanol, which is recrystallised frommethyl alcohol. There is thus obtained the compound Iai, weight 135 g.,M.P. (K): 265 C., [a] =+65.8 (concentration: 1% in 95% alcohol).

EXAMPLE 31 3-dimethy1amino-17a,21-dihydroxy-5- pregnane 11,20- dione(Ia/z) This is prepared according to the procedure of the procedure ofthe previous example, pyrrolidine being replaced by an alcoholicsolution of dimethylamine. No basic product crystallises out directly,and the hydrochloride is directly precipitated from an alcohol solutionof the crude amine. This hydrochloride is then recrystallised toconstant M.P. (K) =265 C.

EXAMPLE 32 21-acetoxy-35-amino-115,17a-dihydroxy-Seregnane- 20-one (lap)and its derivatives There are charged into an apparatus forhydrogenation at ordinary pressure, fitted with an electromagneticstirrer which heats:

4 g. of hydrocortisone acetate (about 0.01 mol) ml. of absolute alcohol,1.2 g. of benzylamine and 0.4 g. of 5% palladium supported uponcharcoal. The mixture is heated to 50 C. and hydrogenation allowed tproceed until 0.03 mol of hydrogen has been absorbed: this requiresabout 8 hours. The mixture is filtered, the residue washed with alcohol,1.5 g. of benzaldehyde is added to the filtrate and the whole is thenheated for 10 minutes under reflux. The alcohol is removed in vacuo, andthe residue taken up in boiling cyclohexane to remove the excess ofaldehyde. The resulting residue is washed with ether and recrystallisedfrom methanol yield of 21- acetoxy-3ebenzylideneimino-l1e,l7a-dihydroxy-5t-pregnane-ZO-one (Iaq), M.P. (tube) 204-205C.,

(concentration 1% in chloroform).

Hydrogenation of the derivative Iaq gives the free amine Iap, ratherunstable and crystallisable with difficulty.

The invention is not limited to the examples the purpose of which is toillustrate and emphasise the great variety of 3-keto steroids and amineswhich can be used.

Thus treatment of 6.3 g. of 5B-pregnane-3,20-dione (IIc) with ammoniaand hydrogen under conditions precisely the same as those used withketone IIb (Example 22, paragraph (b)) gives after recrystallisationfrom ethyl acetate, 0.5 g. of 3p-benzylidene amino-Sfi-pregnane-ZO- one(Is) M.P. (tube) l89l93 C. There is no depression of melting point oncarrying out a mixed melting point'determination with :an authenticspecimen prepared as described in paragraph (a) of Example 21.

In the same way 3 S-dimethylamino-5,8-pregnane-20- one, ormethyl-dihydrohalophylline (Janot and coworkers, Bull. Soc. Chim.France, 1959, page 896) MP. 56 C. (tube), has been prepared by thecatalytic reductive amination of 5u-pregnane-3,20-dione (IIb) anddimethylamine. The fumarate of the base recrystallised from acetone hasM.P. 159161 C. (tube), [u] =+65.9 (concentration=0.8% in methanol).

We claim:

1. A process for the production of 3-a minosteroids having the generalformula H is and R being selected from the group consisting of hydrogenatoms and groups having the formula OR R is selected from the groupconsisting of hydrogen atoms and -OR*, not more than one of R and Rbeing the group --OR R is selected from the group consisting of H OH andR is selected from the group consisting of hydrogen atoms and methylgroups;

X is selected from the group consisting of hydrogen atoms at positions 4and and together an olefinic linkage between positions 4 and 5;

Am is selected from the group consisting of (a) primary amino groups,(b) amino groups containing at least one substituent selected from thegroup consisting of aliphatic groups having less than six carbon atoms,phenylalkyl groups having less than six carbon atoms in the alkylportion thereof, and saturated heterocyclic groups selected from theclass consisting of pyrrolindo, piperidino, morpholino, piperazino andN'-lower alkyl piperazino groups, (0) acetylamido group, and (d) iminogroups having the general formula 30 -N=CHAr in which Ar is selectedfrom the class consisting of phenyl and p-methoxy phenyl, which processcomprises treating a ketone having the general formula ANV in which R isselected from the group consisting of OR and -CR"-CH -R R being selectedfrom the group consisting of hydrogen atoms and acyl groups derived frommonocarboxylic saturated aliphatic acids having less than six carbonatoms, R being selected from the group consisting of =0 and and R beingselected from the group consisting of hydrogen atoms and groups havingthe formula --OR :R is selected from the group consisting of hydrogenatoms and OR not more than one of R and R being the group -OR R isselected from the group consisting of H and =0;

H OH R is selected from the group consisting of hydrogen atoms andmethyl groups;

X is selected from the group consisting of hydrogen atoms at positions 4and 5 and together an olefinic linkage between positions 4 and 5, withfor-mic acid and with a monoamine having the general formula AmH inwhich AM is selected from the group consisting of (a) primary aminogroups, (b) amino groups containing at least one substituent selectedfrom the group consisting of aliphatic groups having less than sixcarbon atoms, phenylalkyl groups having less than six carbon atoms inthe alkyl portion thereof, and saturated heterocyclic groups selectedfrom the class consisting of pyrrolidino, piperidino, morpholino,piperazino and N'-lower alkyl piperazino groups, (c) acetylamido group,and (d) imino groups having the general formula N=CHAr, in which Ar 31is selected from the class consisting of phenyl and pmethoxy phenyl.

3. The process of claim 2, the Xs together defining an olefinic linkagebetween positions 4 and 5 and R representing CO-CH -'R said ketone beingreacted firstly with said monoamine, and subsequently -with formic acid.

4. A process as claimed in claim 1 in which the amine employed is asecondary amine.

5. A process as claimed in claim 2 in which the amine employed is asecondary amine.

6. A process as claimed in claim 1 in which benzylamine is used and thehydrogenation is continued to effect hydrogenolysis of the secondaryamine initially produced thereby to produce a steroid having a primaryamino group in the 3-position.

8. 3fi-amine-21-acetoxy-S/S-pregnane-ZO-one.

9. 3a-pyrrolidinyl-Sfl-pregnane-ZO-one.

10- 318-pyrrolidinyl-5a-pregnane-ZO-One.

11. 3a-pyrrolidinyl-5,8-pregnane-20fi-o1.

12. 3B-pyrrolidinyl-5/3-pregnane-20,8,2l-diol.

13. 3a-piperidinyl-5/8-pregnane-205,2l-dio1.

14. 3a-pyrrolidinyl-5fl-pregnane-20/3,21-diol.

15. 3a-pyrrolidinyl-2'1-hydroxy-5,B-pregnane-20-one.

16. 3BbenZy1idenamino-2 1-acctoXy-5 3pregnane-20-0ne.

0 NM a References Cited UNITED STATES PATENTS 2,884,416 4/1959 Babcoek260-239.5 2,919,285 12/1959 Pappo 260397.3

20 HENRY A. FRENCH, Primary Examiner.

U.S. Cl. X.-R.

UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No.3,424,747

January 28, 1969 Josef Schmitt et a1.

hat error appears in the above identified It is certified t ettersPatent are hereby corrected as patent and that said L shown below:

In the heading to the printed specification, after line 9, insert June17, 1960, 830, 360; June 17 1960, 830,361

Signed and sealed this 14th day of April 1970.

(SEAL) Attest:

WILLIAM E. SCHUYLER, JR.

Edward M. Fletcher, Jr.

Qmnmissioner of Patents Attesting Officer

